Dobretsov Maxim, Petkau Georg, Hayar Abdallah, Petkau Eugen
Department of Anesthesiology, University of Arkansas for Medical Sciences;
Department of Lymphocyte Development, Max Plank Institute for Infection Biology.
J Vis Exp. 2017 Jun 19(124):55819. doi: 10.3791/55819.
The clock scan protocol for image analysis is an efficient tool to quantify the average pixel intensity within, at the border, and outside (background) a closed or segmented convex-shaped region of interest, leading to the generation of an averaged integral radial pixel-intensity profile. This protocol was originally developed in 2006, as a visual basic 6 script, but as such, it had limited distribution. To address this problem and to join similar recent efforts by others, we converted the original clock scan protocol code into two Java-based plugins compatible with NIH-sponsored and freely available image analysis programs like ImageJ or Fiji ImageJ. Furthermore, these plugins have several new functions, further expanding the range of capabilities of the original protocol, such as analysis of multiple regions of interest and image stacks. The latter feature of the program is especially useful in applications in which it is important to determine changes related to time and location. Thus, the clock scan analysis of stacks of biological images may potentially be applied to spreading of Na or Ca within a single cell, as well as to the analysis of spreading activity (e.g., Ca waves) in populations of synaptically-connected or gap junction-coupled cells. Here, we describe these new clock scan plugins and show some examples of their applications in image analysis.
用于图像分析的时钟扫描协议是一种有效的工具，可量化封闭或分割的凸形感兴趣区域内、边界处及外部（背景）的平均像素强度，从而生成平均积分径向像素强度剖面图。该协议最初于2006年作为Visual Basic 6脚本开发，但当时其传播有限。为了解决这个问题并加入其他人最近的类似努力，我们将原始的时钟扫描协议代码转换为两个基于Java的插件，这些插件与美国国立卫生研究院赞助的、可免费获取的图像分析程序（如图像J或Fiji ImageJ）兼容。此外，这些插件还有几个新功能，进一步扩展了原始协议的功能范围，如对多个感兴趣区域和图像堆栈的分析。该程序的后一个功能在确定与时间和位置相关的变化很重要的应用中特别有用。因此，生物图像堆栈的时钟扫描分析可能潜在地应用于单个细胞内钠或钙的扩散，以及对突触连接或间隙连接耦合细胞群体中扩散活动（如钙波）的分析。在这里，我们描述这些新的时钟扫描插件，并展示它们在图像分析中的一些应用示例。
